In previous studies we have described the dispersion by detergents of adenylate cyclase from brain and from partially purified plasma membranes from liver. The detergent-dispersed enzymes have been characterized with respect to stability, effects of fluoride, cations, and chelators. In addition, we have achieved some success in partially purifying the enzyme, particularly with ion exchange an hydrophobic chromatography. Our objectives in this proposal are as follows: 1) To purify adenylate cyclase. We will initially use detergent-dispersed enzyme with ion exchange chromotography and gel filtration techniques, but plan to evaluate more completely the potential usefulness of hydrophobic chromatography. In addition we plan to develop an affinity chromatography system for the brain cyclase by coupling EGTA to agarose. 2) To establish if there is a causal relationship between membrane phosphorylation and alterations in cyclase activity caused by F minus or glucagon. We will determine if there are membrane-bound phosphatase and kinase which would regulate adenylate cyclase. These studies will be done in conjunction with the purification of the enzyme. 3) Eventually, to reconstitute the adenylate cyclase system, which comprises hormone receptor, the catalytic moiety, and other factor(s) associated with the control of adenylate cyclase.